Photoactivatable RNA N6 -Methyladenosine Editing with CRISPR-Cas13.
Photoactivatable RNA N6 -Methyladenosine Editing with CRISPR-Cas13.
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DOI：
10.1002/smll.201907301
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发表时间：
2020-06
期刊：
影响因子：
13.3
通讯作者：
Jie Zhao;Bing Li;Jian-xiong Ma;Weilin Jin;Xinlong Ma
中科院分区：
文献类型：
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作者：
Jie Zhao;Bing Li;Jian-xiong Ma;Weilin Jin;Xinlong Ma
RNA具有重要而多样的生物学作用，但时空调控的分子方法有限。在这里，设计了一种具有CRISPR-Cas 13的工程化光活化RNA N6 -甲基腺苷（m6 A）编辑系统，以指导特异性m6 A编辑。光诱导异源二聚化蛋白CIBN和CIB 2 PHR分别与无催化活性的PguCas 13（dCas 13）和m6 A效应子融合。该系统被称为PAMEC，能够响应于蓝光对m6 A编辑进行时空控制。进一步优化该系统以创建高效版本，称为PAMEC R，允许同时稳健地操纵多个基因。当与上转换纳米颗粒膜结合时，光遗传学操作窗口从可见光范围扩展到组织可穿透的近红外波长，这为远程控制RNA编辑提供了一个有吸引力的途径。这些结果表明，PAMEC是一种有前途的光遗传学平台，可灵活有效地靶向RNA，对表观转录组工程、成像和未来治疗开发具有广泛的适用性。
RNA has important and diverse biological roles, but the molecular methods to manipulate it spatiotemporally are limited. Here, an engineered photoactivatable RNA N6 -methyladenosine (m6 A) editing system with CRISPR-Cas13 is designed to direct specific m6 A editing. Light-inducible heterodimerizing proteins CIBN and CRY2PHR are fused to catalytically inactive PguCas13 (dCas13) and m6 A effectors, respectively. This system, referred to as PAMEC, enables the spatiotemporal control of m6 A editing in response to blue light. Further optimization of this system to create a highly efficient version, known as PAMECR , allows the manipulation of multiple genes robustly and simultaneously. When coupled with an upconversion nanoparticle film, the optogenetic operation window is extended from the visible range to tissue-penetrable near-infrared wavelengths, which offers an appealing avenue to remotely control RNA editing. These results show that PAMEC is a promising optogenetic platform for flexible and efficient targeting of RNA, with broad applicability for epitranscriptome engineering, imaging, and future therapeutic development.